LOCUS       sequence-301939-        8616 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of SARS-CoV-2 S protein (D614) with the flag 
            tag at N- and C-termini. Pseudotypes onto the MLV vector but not 
            efficiently to VSV or lentiviral vectors..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8616)
  AUTHORS   Zhang L, Jackson CB, Mou H, Ojha A, Peng H, Quinlan BD, Rangarajan 
            ES, Pan A, Vanderheiden A, Suthar MS, Li W, Izard T, Rader C, Farzan
            M, Choe H
  TITLE     SARS-CoV-2 spike-protein D614G mutation increases virion spike 
            density and infectivity.
  JOURNAL   Nat Commun. 2020 Nov 26;11(1):6013. doi: 10.1038/s41467-020-19808-4.
  PUBMED    33243994
REFERENCE   2  (bases 1 to 8616)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2020-11-26"; volume: "11(1):6013. doi"; pages: " 
            10.1038/s41467-020-19808-4"
COMMENT     Sequence Label: pCAGGS-SARS2-S-G614 (NC-flag)
FEATURES             Location/Qualifiers
     source          1..8616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1674
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1682..1701
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1783..1806
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5587..5610
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(5638..5657)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5703..5758
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5757..5776)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6119..6135)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6119..6135)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6132..6154)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6143..6159
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6167..6173,6174..6191,6192..6197))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6212..6233
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6292..6487
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6338..6473
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6493..6627
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6530..6549)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6584..6603
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6695..6712)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6866..7454)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6946..6965)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7625..8416,8417..8485))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8248..8267
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8486..8590)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1681 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattcgc cgccaccatg
     1741 tttgtgttcc tggtcctgct cccactggtt tccagccaat gtgattacaa ggatgacgat
     1801 gacaaggtta acctcacaac ccgcacccag ctgcctcccg cctataccaa cagttttact
     1861 cggggagtgt attaccctga caaagttttc aggagctcag ttctgcactc aacccaggat
     1921 ctctttctgc cttttttttc caacgtgact tggttccatg caattcacgt ttccgggacc
     1981 aacggcacca aaaggttcga taaccccgtg ctgcccttta acgacggcgt ctatttcgca
     2041 agcactgaga aaagcaatat catcaggggc tggatttttg ggacaacctt ggatagcaag
     2101 acccagtcac tgctgatcgt taataatgca accaatgtgg tgatcaaagt gtgcgaattt
     2161 cagttttgca atgacccctt tctgggcgtc tactatcata aaaacaataa atcttggatg
     2221 gagtctgagt ttagggtgta ttctagcgcg aataattgta cctttgaata tgtatcacaa
     2281 ccatttctca tggatctcga aggcaaacaa ggcaatttca aaaacttgag agagttcgtc
     2341 ttcaaaaaca tcgatggata ttttaaaata tacagcaaac ataccccaat aaacctggtc
     2401 cgcgacctcc ctcagggttt cagcgccctc gagcctctcg tcgatcttcc tatcgggatc
     2461 aacatcacaa gatttcaaac cttgctggct cttcatcgat cctacctgac ccctggcgat
     2521 tcctcttccg ggtggaccgc cggcgccgct gcctattatg tggggtatct gcaacctcgc
     2581 acgtttctcc tcaagtataa tgagaatgga actatcaccg atgcggtaga ctgtgcgttg
     2641 gacccactct ctgagaccaa gtgcactttg aagtccttta ccgttgaaaa ggggatctat
     2701 cagacatcca attttagagt ccaacccacg gaatccatcg ttcggttccc caatatcaca
     2761 aatctctgcc ctttcggaga ggtattcaac gcaactcgct tcgcttctgt gtatgcgtgg
     2821 aacagaaaac gcatctccaa ctgcgtcgct gattactctg ttctgtataa ctccgcctcc
     2881 tttagcacct tcaagtgtta cggcgtcagc cccacaaagc tcaacgactt gtgtttcact
     2941 aacgtctacg ccgactcctt cgtgatccgg ggggacgaag ttcggcagat tgcgcctgga
     3001 cagactggta aaatcgctga ctataactat aagctcccag atgactttac cggctgtgtg
     3061 attgcttgga actctaataa tctcgacagc aaagttggag gcaactataa ttatctctac
     3121 cggttgtttc ggaagagcaa tctcaagcca ttcgagagag acataagcac ggagatctac
     3181 caagccggct ccactccatg caatggggtt gaaggcttta attgttattt tcccctccaa
     3241 tcctatggtt ttcaaccgac taatggagtc gggtatcaac cctacagggt tgtcgtcttg
     3301 agcttcgagc tcttgcacgc tcctgctacg gtgtgtggtc cgaaaaaatc aacaaacctc
     3361 gtcaaaaaca aatgtgtgaa ttttaacttt aacggactga ctggcaccgg ggtgctgact
     3421 gagagcaata agaagtttct tccatttcaa caattcggtc gggatatcgc ggacaccact
     3481 gatgctgtgc gcgatccgca aaccttggag atcttggaca taactccgtg cagcttcgga
     3541 ggcgtctctg ttatcacccc tggaacgaat acatctaacc aagttgcggt cctttatcag
     3601 ggtgttaatt gcacagaagt acccgtcgcg atacatgcgg atcagctgac ccctacttgg
     3661 cgggtctact caaccggctc taatgtgttt caaactcggg cgggatgttt gatcggagct
     3721 gagcacgtga ataactctta tgaatgcgat ataccaatcg gcgcagggat ttgcgcatct
     3781 taccaaaccc aaaccaactc tcctcggcgg gctcgctccg tcgcatccca atccattatc
     3841 gcctatacca tgagtctcgg agctgagaac tctgtcgctt acagcaacaa ttctatagcc
     3901 atcccaacca acttcacgat tagcgttaca acagagatac tccccgtcag catgacgaaa
     3961 acttccgttg attgtacgat gtatatctgt ggggattcta cagagtgtag caatctgctg
     4021 ttgcagtatg gctccttctg tacccaattg aatcgggcac tcaccggcat cgctgtggaa
     4081 caagacaaaa atacgcaaga agttttcgca caggtgaagc aaatctacaa gacgcctcca
     4141 atcaaagact ttggcggatt caatttttct cagattctgc ctgatccctc aaagcccagc
     4201 aaaagaagct tcatcgagga cctgctcttc aacaaggtca ctctcgcaga tgccgggttc
     4261 atcaaacaat atggagattg cttgggcgac attgctgcgc gggatcttat ttgtgctcaa
     4321 aagttcaatg ggctgaccgt cctgccgccc ctcttgaccg atgaaatgat tgctcaatac
     4381 acctccgcgt tgctcgcggg aactatcact tccggctgga ccttcggcgc cggcgctgcc
     4441 ctccagatac cgtttgccat gcaaatggcc tacagattca acggcatcgg agtcacacag
     4501 aatgtcctgt acgaaaatca aaaattgata gccaatcagt ttaattctgc aatcggcaaa
     4561 atacaagact ccctgtcctc tactgcctcc gcacttggga aattgcaaga cgtcgtgaac
     4621 caaaacgcac aggcccttaa cactctggtg aaacagttga gctccaactt cggcgctata
     4681 agctccgtct tgaacgatat actctctcgc ctggacaaag tcgaggctga agtccagatc
     4741 gacagattga taacaggcag gcttcaatcc ctgcaaacct acgtcacgca gcagctgatc
     4801 agagccgcag agatcagggc ttccgcaaat cttgctgcaa caaaaatgtc cgaatgcgtc
     4861 ttgggccaat caaagcgcgt cgatttctgt ggaaagggct accatctcat gagctttccc
     4921 caaagcgccc cgcacggcgt ggtttttctc cacgtcacct acgtccccgc tcaagagaag
     4981 aactttacaa ccgctcctgc aatttgccat gatgggaaag cgcactttcc tcgcgaaggc
     5041 gtctttgtct ccaatggcac acattggttt gttactcaac ggaactttta tgaaccacaa
     5101 atcattacca cggacaacac gttcgtgtcc ggcaactgtg acgttgtcat cggaattgtt
     5161 aacaacacgg tttacgatcc tctgcaaccc gagcttgact cctttaagga agaactggac
     5221 aagtacttca aaaatcacac ctcaccggat gtggacttgg gcgatatctc tggcataaat
     5281 gcgtcagtcg tgaacattca aaaggaaatt gatcgactca acgaggtcgc aaagaatctt
     5341 aatgaatccc tgatagatct gcaggagttg gggaagtacg agcagtatat aaagtggcct
     5401 tggtatatat ggcttggctt tatcgccggc ctcattgcaa tcgtgatggt caccatcatg
     5461 ctgtgctgca tgacctcatg ttgctcctgt cttaaggggt gctgctcctg cggttcctgt
     5521 tgcaaatttg acgaggacga cagcgaaccg gtgcttaagg gtgtgaagct gcattacacc
     5581 ggaggcgatt acaaggatga cgatgacaag tgacggccgc ctgcaggtta attaagatct
     5641 ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg
     5701 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
     5761 cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag
     5821 tttggcaaca tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca
     5881 tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg
     5941 aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt
     6001 ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc
     6061 tgtccctctt ctcttatgga gatccctcga cctgcagccc aagcttggcg taatcatggt
     6121 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6181 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6241 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa
     6301 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     6361 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     6421 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     6481 ttgcaaaaag ctaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     6541 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     6601 atcaatgtat cttatcatgt ctggatccgc tgcattaatg aatcggccaa cgcgcgggga
     6661 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6721 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6781 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6841 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6901 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6961 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     7021 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
     7081 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     7141 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     7201 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     7261 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     7321 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     7381 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     7441 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     7501 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7561 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7621 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7681 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7741 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     7801 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7861 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     7921 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7981 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     8041 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     8101 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     8161 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     8221 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     8281 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     8341 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     8401 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     8461 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     8521 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8581 gggttccgcg cacatttccc cgaaaagtgc cacctg
//
